How can I quantitively analyze ELISA?

05 June 2016 4,074 3 View

Does anyone have experience with S.mutans UA159?

I do transformations of erythromycin resistant plasmids into S.mutans. I am short in time. Do you know what is the working concentration of S.mutans UA159 to erythromycin? I tried with 10 ug/ml,...

08 September 2015 9,145 1 View

Is it possible to transform pet21 vector into gram-positive bacteria successfully?

07 August 2015 8,961 2 View

Which of the Covid-19 Vaccines would elicit the best immunoprotection and why?

I want to know which of the available Covid-19 Vaccines would elicit the best immunological response and why

02 March 2021 1,388 3 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

Experimental Design: How deal with pseudreplicates with immunological and clinical-chemical paramaeters?

Hello everybody, I am investigating the influence of a toxinon various clinical-chemical and haematological parameters in fish. I have three treatments (control, low dose, high dose) in my...

24 February 2021 1,243 1 View

Looking for advices and tips to improve the staining and the gating strategy in human blood?

Hi, I am staining a human blood with CD19, CD3,CD14 and CD16. My gating strategy in FlowJo goes like this: - First, I gate a general population SSC-A VS FSC-A => Then gate for single cells, =>...

21 February 2021 5,724 4 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View

How can I completely delete a protein on the genomic level using Crispr?

I want to completely delete or largely truncate a gene and not only achieve a functional knockout in a cell line. I can only transfect my cells with the Crispr/Cas9 sgRNA RNP and not introduce a...

08 February 2021 2,032 2 View

Does gene insertion via CRISPR/Cas9 result in chromosomal abnormalities?

Hello, I hope to insert a gene into mammalian cells in culture using CRISPR/Cas9 and an HDR template. Will insertion of a 1 kilobase gene cause chromosomal abnormalities and effect cell...

31 January 2021 2,516 1 View

Should I add a promoter upstream of my CRISPR-inserted gene?

Hello, I want to insert a gene into my cells using sgRNA, Cas9, and an HDR template. The gene that I will insert into has its own promoter. Because I will cut the gene downstream of its start...

14 January 2021 1,170 2 View

Can you polarize Jurkat cells into different T helper subsets?

I am wondering if anyone has had success with in vitro polarization (via cytokines) of Jurkat cells. My goal is to understand the affect of a drug on Treg induction and differentiation. I...

12 January 2021 3,606 2 View

Does anybody have experience in expressing and incorporating two proteins (to express as a protein complex) in one cloning site?

I want to clone have two proteins cloned into pPROexHtb+ (which has one cloning site). I want to express them as a protein complex. My question is would you generally put a stop codon after first...

09 January 2021 8,811 2 View